PARENT SESSION
IVF and Assisted Reproductive Technologies

CAPRINE IN VITRO FERTILIZATION UTILIZING CRYOPRESERVED EPIDIDYMAL OR EJACULATE TRANSGENIC SPERM.

Melican, David¹, Blash, Stephen¹, Gavin, William¹, ¹

ABSTRACT- The ability to successfully collect and cryopreserve both epididymal and ejaculated sperm from a transgenic founder male dairy goat for progeny development is paramount. The goal of this experiment was to determine the feasibility of using both cryopreserved epididymal or ejaculated sperm from a transgenic buck for use in an IVF system. One hundred and seven ovulated oocytes and 127 aspirated COCs with intact and expanded cumulus were recovered from thirty superovulated does out-of-season (August). Two separate IVF experiments were performed utilizing percoll-purified cryopreserved epididymal or ejaculated sperm from a single two-year old transgenic F2 male. IVF was performed in B-O media supplemented with 20% heat-inactivated estrus goat serum, 7.7 mM calcium lactate, 100 U/ml penicillin and 100 mcg/ml streptomycin for 18 hours. Following IVF, presumptive zygotes were transferred to equilibrated SOF/BSA for 24 hours. On Day 2 cleavage was determined, and then five 2 to 8-cell embryos produced from each sperm type were surgically transferred to the oviducts of seven synchronized recipients. The remaining embryos were allowed to continue development in vitro to Day 7 to 9, then two or three blastocysts produced from each sperm type were surgically transferred to the uterine horn of five or six synchronized recipients. Pregnancy was determined by ultrasound on Days 35 and 50. Overall cleavage (67% with epididymal sperm versus 57% with ejaculated sperm) and blastocyst development (21% with epididymal sperm versus 17% with ejaculated sperm) was comparable. Pregnancy rates for Day 2 transfers were equivalent (57% for either epididymal or ejaculated sperm), while only blastocysts produced with epididymal sperm resulted in pregnancies. In conclusion, IVF performed utilizing cryopreserved epididymal or ejaculated transgenic sperm resulted in successful cleavage, blastocyst development and pregnancies, and offers a potential alternative method for progeny development of production animals from transgenic founder male dairy goats.

KEY WORDS: caprine, transgenic, epididymal , IVF